Immunofluorescence staining of GenCRISPRᵀᴹ SpCas9 Antibody (4A1) (GenScript, A01935-40) with SpCas9 in Hela cells.
HeLa cells transfected with PX458 (pSpCas9(BB)-2A-GFP) (green) were fixed with 4% Poly-Formaldehyde (5min) and then blocked in 3% BSA 30min.
The cells were then incubated with GenCRISPRᵀᴹ SpCas9 Antibody (4A1) (GenScript, A01935-40) at 0.25 μg/ml, 0.5 μg/ml, 1 μg/ml and mouse IgG at 1 μg/ml at room temperature for 2h, followed by a further incubation at 37℃ for 1h with Goat Anti-Mouse IgG Antibody (H&L) [ifluor 555], pAb (GenScript) (red) at 5 μg/ml.
DAPI was used to stain the cell nuclei (blue) at a concentration of 0.2 μg/ml for 2h at room temperature.

Standard curve of SpCas9 Sandwich ELISA. The SpCas9 Sandwich ELISA assay is developed by using GenCRISPRᵀᴹ SpCas9 Antibody (14B6) (GenScript, A01936-40) and GenCRISPRᵀᴹ SpCas9 Antibody (4A1) (GenScript, A01935-40) as capture and detection antibody, respectively. These two antibodies recognize different epitopes.
In this ELISA assay, GenCRISPRᵀᴹ SpCas9 Antibody (4A1) (GenScript, A01935-40) was labeled with Biotin. GenScript can provide customized conjugation service for this product per customer's request.
The sensitivity is <1 ng/ml and the detection range is 0-10 ng/ml.

Clustered regularly interspaced short palindromic repeats (CRISPR)/CRISPR-associated (Cas) protein 9 system provides a robust and multiplexable genome editing tool, enabling researchers to precisely manipulate specific genomic elements, and facilitating the elucidation of target gene function in biology and diseases. CRISPR/Cas9 genome editing allows for double-stranded DNA breaks at specific sequences to efficiently disrupt, excise, mutate, insert, or replace genes. To date, the Streptococcus pyogenes Cas9 (SpCas9) has been used broadly to achieve efficient genome editing in a variety of species and cell types. It is important that the precision of transfection and the level of Cas9 expression should be controlled strictly during the editing processes by using specific anti-CRISPR/Cas9 antibodies.

Reconstitute the lyophilized antibody with deionized water (or equivalent) to a final concentration of 0.5 mg/ml.

Specificity

The product is specific for Streptococcus pyogenes CRISPR/Cas9. This antibody binds with recombinant Streptococcus pyogenes CRISPR/Cas9 protein in ELISA and Endogenous overexpressed Streptococcus pyogenes CRISPR/Cas9 in immunofluorescence, western blot.

Storage

The lyophilized product remains stable up to 1 year at -20 °C from date of receipt. Upon reconstitution, it can be stored for 2-3 weeks at 2-8 °C or for up to 12 months at -20 °C or below. Avoid freeze/thaw cycles.

Working concentrations for specific applications should be determined by the investigators. The appropriate concentration may be affected by secondary antibody affinity, antigen concentration, the sensitivity of the method of detection, temperature, the length of the incubations, and other factors. The suitability of this antibody for applications other than those listed below has not been determined. The following concentration ranges are recommended starting points for this product.

Immunofluorescence staining of GenCRISPRᵀᴹ SpCas9 Antibody (4A1) (GenScript, A01935-40) with SpCas9 in Hela cells.
HeLa cells transfected with PX458 (pSpCas9(BB)-2A-GFP) (green) were fixed with 4% Poly-Formaldehyde (5min) and then blocked in 3% BSA 30min.
The cells were then incubated with GenCRISPRᵀᴹ SpCas9 Antibody (4A1) (GenScript, A01935-40) at 0.25 μg/ml, 0.5 μg/ml, 1 μg/ml and mouse IgG at 1 μg/ml at room temperature for 2h, followed by a further incubation at 37℃ for 1h with Goat Anti-Mouse IgG Antibody (H&L) [ifluor 555], pAb (GenScript) (red) at 5 μg/ml.
DAPI was used to stain the cell nuclei (blue) at a concentration of 0.2 μg/ml for 2h at room temperature.

Standard curve of SpCas9 Sandwich ELISA. The SpCas9 Sandwich ELISA assay is developed by using GenCRISPRᵀᴹ SpCas9 Antibody (14B6) (GenScript, A01936-40) and GenCRISPRᵀᴹ SpCas9 Antibody (4A1) (GenScript, A01935-40) as capture and detection antibody, respectively. These two antibodies recognize different epitopes.
In this ELISA assay, GenCRISPRᵀᴹ SpCas9 Antibody (4A1) (GenScript, A01935-40) was labeled with Biotin. GenScript can provide customized conjugation service for this product per customer's request.
The sensitivity is <1 ng/ml and the detection range is 0-10 ng/ml.